AmpliTaq and AmpliTaq Gold DNA Polymerase - Applied Biosystems

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and MSP2 protein levels per A. marginale organism increase only minimally and transiently insalivary glands of transmission-feeding ticks compared to that of unfed ticks. A. marginalenumbers per tick increase gradually in salivary glands of both transmission-fed and unfed ticks. Itis concluded that MSP2 variant selection is an early event in the tick and that MSP2 variantsSGV1 and SGV2 are expressed both in the midgut and salivary glands. While MSP2 may berequired for infectivity, there is no strict temporal correlation between MSP2 expression and thedevelopment of infectivity.Mederle, I., I. Bourguin, et al. (2002). "Plasmidic versus Insertional Cloning of Heterologous Genes inMycobacterium bovis BCG: Impact on In Vivo Antigen Persistence and Immune Responses."Infect. Immun. 70(1): 303-314.http://iai.asm.org/cgi/content/abstract/70/1/303Bivalent recombinant strains of Mycobacterium bovis BCG (rBCG) expressing the early regulatorynef and the structural gag(p26) genes from the simian immunodeficiency virus (SIV) SIVmac251were engineered so that both genes were cotranscribed from a synthetic operon. The expressioncassette was cloned into a multicopy-replicating vector, and the expression levels of both nef andgag in the bivalent rBCG(nef-gag) strain were found to be comparable to those of monovalentrBCG(nef) or rBCG(gag) strains. However, extrachromosomal cloning of the nef-gag operon intoa replicative plasmid resulted in strains of low genetic stability that rapidly lost the plasmid in vivo.Thus, the nef-gag operon was inserted site specifically into the BCG chromosome by means ofmycobacteriophage Ms6-derived vectors. The resulting integrative rBCG(nef-gag) strains showedvery high genetic stability both in vitro and in vivo. The in vivo expression of the heterologousgenes was much longer lived when the expression cassette was inserted into the BCGchromosome. In one of the strains obtained, integrative cloning did not reduce the expressionlevels of the genes even though a single copy was present. Accordingly, this strain inducedcellular immune responses of the same magnitude as that of the replicative rBCG straincontaining several copies of the genes.Reglier-Poupet, H., E. Pellegrini, et al. (2003). "Identification of LpeA, a PsaA-Like Membrane ProteinThat Promotes Cell Entry by Listeria monocytogenes." Infect. Immun. 71(1): 474-482.http://iai.asm.org/cgi/content/abstract/71/1/474The intracellular life of Listeria monocytogenes starts by a complex process of entry involvingseveral bacterial ligands and eukaryotic receptors. In this work, we identified in silico from thesequence of the genome of L. monocytogenes a previously unknown gene designated lpeA (forlipoprotein promoting entry) encoding a 35-kDa protein homologous to PsaA, a lipoproteinbelonging to the LraI family and implicated in the cell adherence of Streptococcus pneumoniaeand related species. By constructing a mutant of L. monocytogenes in which lpeA is deleted (lpeAmutant), we show that the PsaA-like protein LpeA is not involved in bacterial adherence but isrequired for entry of L. monocytogenes in eukaryotic cells. In contrast to wild-type bacteria,mutant bacteria failed to invade the epithelial Caco-2 and hepatocyte TIB73 cell lines, asconfirmed by confocal microscopy. The mutant bacteria rapidly penetrated in mouse bonemarrow-derived macrophages. Surprisingly, lpeA mutant bacteria survive better in macrophagesthan do wild-type bacteria. This was correlated with a weak exacerbation of virulence of the lpeAmutant in the mouse. LpeA is therefore a novel invasin favoring the entry of L. monocytogenesinto nonprofessional phagocytes but not its invasion of macrophages. This is the first report of alipoprotein promoting cell invasion of an intracellular pathogen.
Judge, N. A., H. S. Mason, et al. (2004). "Plant Cell-Based Intimin Vaccine Given Orally to Mice Primedwith Intimin Reduces Time of Escherichia coli O157:H7 Shedding in Feces." Infect. Immun. 72(1):168-175.http://iai.asm.org/cgi/content/abstract/72/1/168Intimin is the primary adhesin of Escherichia coli O157:H7, the most common infectious cause ofbloody diarrhea in the United States and the leading cause of acute kidney failure in children whodevelop hemolytic uremic syndrome. Cattle are the primary reservoir of E. coli O157:H7. Indeed,most cases of E. coli O157:H7 infection in the United States occur after ingestion of contaminatedundercooked hamburger or produce that had contact with bovine manure. Because intimin isrequired for persistent colonization of neonatal calves and adult cattle, we hypothesized that anintimin-based vaccination strategy in calves would reduce colonization of cattle with E. coliO157:H7. To test this concept in a small-animal model, we developed transgenic tobacco plantcells that express the carboxy-terminal host cell-binding domain of E. coli O157:H7 intimin. Micewere either immunized intraperitoneally with intimin expressed from the plant cells, fed transgenicplant cells, or both. Here we show that these mice generated an intimin-specific mucosal immuneresponse when primed parenterally and then boosted orally and also exhibited a reducedduration of E. coli O157:H7 fecal shedding after challenge.Munoz-Elias, E. J., J. Timm, et al. (2005). "Replication Dynamics of Mycobacterium tuberculosis inChronically Infected Mice." Infect. Immun. 73(1): 546-551.http://iai.asm.org/cgi/content/abstract/73/1/546The dynamics of host-pathogen interactions have important implications for the design of newantimicrobial agents to treat chronic infections such as tuberculosis (TB), which is notoriouslyrefractory to conventional drug therapy. In the mouse model of TB, an acute phase of exponentialbacterial growth in the lungs is followed by a chronic phase characterized by relatively stablenumbers of bacteria. This equilibrium could be static, with little ongoing replication, or dynamic,with continuous bacterial multiplication balanced by bacterial killing. A static model predicts aclose correspondence between "viable counts" (live bacteria) and "total counts" (live plus deadbacteria) in the lungs over time. A dynamic model predicts the divergence of total counts andviable counts over time due to the accumulation of dead bacteria. Here, viable counts are definedas bacterial CFU enumerated by plating lung homogenates; total counts are defined as bacterialchromosome equivalents (CEQ) enumerated by using quantitative real-time PCR. We show thatthe viable and total bacterial counts in the lungs of chronically infected mice do not diverge overtime. Rapid degradation of dead bacteria is unlikely to account for the stability of bacterial CEQnumbers in the lungs over time, because treatment of mice with isoniazid for 8 weeks led to amarked reduction in the number of CFU without reducing the number of CEQ. Theseobservations support the hypothesis that the stable number of bacterial CFU in the lungs duringchronic infection represents a static equilibrium between host and pathogen.De Gregorio, E., C. Abrescia, et al. (2003). "Asymmetrical Distribution of Neisseria Miniature InsertionSequence DNA Repeats among Pathogenic and Nonpathogenic Neisseria Strains." Infect.Immun. 71(7): 4217-4221.http://iai.asm.org/cgi/content/abstract/71/7/4217
Page 8 and 9: succinyltransferase. Antiserum from
Page 10 and 11: inoculated with N40 by syringe than
Page 12 and 13: Post, D. M. B., N. J. Phillips, et
Page 14 and 15: 700.http://iai.asm.org/cgi/content/
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Page 20 and 21: Behrens, M., J. Sheikh, et al. (200
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Page 25 and 26: histopathology. The mucosal immune
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Page 29 and 30: Immun. 70(10): 5556-5561.http://iai

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